Abstract:

BACKGROUND: Tissue engineering requires a lot of seed cells. Osteoblasts have become an important seed cells in bone tissue engineering. However, there are the difficulties to derive osteoblasts and the different purity of osteoblasts was obtained.
OBJECTIVE: To establish the methods of isolated culture and purification of neonatal rat calvarial osteoblasts, and to observe the biological characteristics of the osteoblasts from the skull.
METHODS: Osteoblasts of Sprague-Dawley neonatal rats were primarily cultured and proliferated by the second enzyme digestion. Osteoblasts were purified by differential attachment method. Osteoblast proliferation and osteogenic activity were identified by morphology, alkaline phosphatase detection, Alizarin red staining and calcium nodules Von kossa staining, ultrastructure and cell proliferation curve.
RESULTS AND CONCLUSION: Primarily cultured calvarial osteoblasts could proliferate by the secondary source of enzyme digestion. The amplification cells showed representative morphological and biological characteristics of osteoblasts. Alkaline phosphatase, Alizarin red staining and calcium nodules Von kossa staining presented positive results. Ultrastructural features appeared as high-differentiated active osteoblasts. Cell proliferation curves showed active cells. Results indicated that calvarial osteoblast of Sprague-Dawley rat has good proliferation and osteogenic activity and can be continuously passaged, with high purity, cell biological characteristics of stability. Calvarial osteoblast is suitable for experiment in vitro.